Manipulating cholangiocarcinoma immune-phenotype in a patient derived precisioncut tumour model to improve immune checkpoint inhibition response.

胆管癌（CCA）是一种侵袭性癌症，其发病率和死亡率在全球范围内呈上升趋势。患者预后仍然很差，免疫治疗方法，如免疫检查点阻断（ICB）与常规化疗相结合，仅在一小部分患者中显示出显着的益处。为了更好地理解治疗反应/耐药机制，有必要进一步研究CCA中的肿瘤微环境（TME）。icb介导的抗肿瘤反应依赖于细胞毒性t细胞的功能来靶向和破坏肿瘤细胞。免疫“热”肿瘤增加了CD8+ t细胞浸润，从而改善了ICB治疗反应，而免疫“冷”肿瘤显示细胞毒性t细胞浸润减少，具有高水平的免疫抑制免疫细胞（例如肿瘤相关巨噬细胞和t调节细胞）。对ICB治疗的不良反应表明许多CCA肿瘤具有“冷”免疫表型。调节肿瘤免疫表型从“冷”到“热”的组合策略可以显着提高ICB的临床有效性，并且仍然是CCA研究中未充分探索的领域。本项目的主要目的是评估人类患者来源的CCA精确切割组织切片（hPCTS）作为离体模型，以研究增加TME的方法，确定免疫治疗的反应/耐药性，并测试新的联合疗法。该项目的第一个目标将是通过药理抑制免疫抑制细胞类型（如肿瘤相关巨噬细胞和髓源性抑制细胞），增强CCA TME并在CCA hPCTS中建立免疫“热”表型。下一步将是确定免疫“热”PCTS TME是否会增加ICB反应，方法是将免疫“热”和控制的hPCTS暴露于ICB抑制剂中，作为单一疗法或与常规化疗联合使用，并评估它们的生存能力。最后，将通过原位数字空间分析建立对ICB反应/耐药性的免疫蛋白质组学预测生物标志物。总的来说，该项目旨在增加CCA hPCTS TME，并评估对各种免疫治疗/化疗方案的反应/耐药性，这有可能改善CCA患者的ICB反应。同时，这项工作将进一步探索免疫治疗反应/耐药机制和识别相应的预测性生物标志物。希望这个模型也将最终取代免疫肿瘤学领域对动物实验的需求，这些实验昂贵，引起伦理问题，并且无法完全概括人类TME和人类疾病。

Cholangiocarcinoma (CCA) is an aggressive cancer type, and its incidence and mortality are increasing globally. Patient prognosis remains poor, with immunotherapeutic approaches such as immune-checkpoint blockade (ICB) in combination with conventional chemotherapy showing a significant benefit in only a small proportion of patients. Further work looking into the tumour microenvironment (TME) in CCA is necessary in order to better understand the mechanisms of therapy response/ resistance. ICB-mediated anti-tumour responses rely on the function of cytotoxic T-cells to target and destroy tumour cells. Immune 'hot' tumours have increased CD8+T-cell infiltration and, thus, improved ICB therapy response, whereas, immune 'cold' tumours show reduced cytotoxic T-cell infiltration and have high levels of immunosuppressive immune cells (e.g. tumour-associated macrophages and T-regulatory cells). The poor response to ICB therapy suggests many CCA tumours have a 'cold' immune phenotype. Combination strategies that modulate tumour immune-phenotype from 'cold' to 'hot' could significantly increase ICB clinical-effectiveness and remains an underexplored area in CCA research. The main aim of this project is to evaluate human patient-derived CCA precision-cut tissue slices (hPCTS) as an ex-vivo model to investigate approaches to augment the TME, determine response/ resistance to immunotherapy and test novel combination therapies. The first objective of the project will be to augment the CCA TME and establish an immune 'hot' phenotype in CCA hPCTS through pharmacological inhibition of immunosuppressive cell types such as tumour-associated macrophages and myeloid-derived suppressor cells. The next step will be to determine whether an immune 'hot' PCTS TME increases ICB response by exposing immune 'hot' and control hPCTS to ICB inhibitors as monotherapy or in combination with conventional chemotherapy and assessing their viability. Finally, immunoproteomic predictive biomarkers of response/ resistance to ICB will be established through in situ digital spatial profiling. Overall, this project aims to augment the CCA hPCTS TME and assess responses/ resistance to various immunotherapeutic/ chemotherapeutic regimes, which has potential to improve ICB response in CCA patients. Simultaneously, this work will further the exploration of immunotherapy response/ resistance mechanisms and the identification of corresponding predictive biomarkers. The hope is that this model will also eventually replace the need for animal experiments in this field of immuno-oncology, which are expensive, raise ethical concerns, and are unable to fully recapitulate the human TME and human disease.